CHD5 CRISPR Activation Plasmid (h): sc-402429-ACT

CHD5 (chromodomain helicase DNA-binding protein 5) is a neuron-enriched ATP-dependent chromatin remodeler in the SNF2 family that coordinates nucleosome positioning and transcriptional programs required for differentiation and genome stability. Through its chromodomains and helicase/ATPase activity, CHD5 modulates chromatin accessibility at regulatory elements and interfaces with epigenetic pathways controlling cell-cycle progression, DNA damage responses, and lineage-specific gene expression. Reduced CHD5 activity has been associated with disrupted transcriptional control and chromosomal instability, and altered CHD5 expression is frequently linked to tumor-suppressive networks in neuroblastoma and other malignancies. In biomedical research, CHD5 is widely studied for its roles in chromatin dynamics, neuronal development, and mechanisms by which epigenetic dysregulation contributes to disease-relevant phenotypes.

CHD5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CHD5 expression without altering the underlying DNA sequence.

CHD5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CHD5 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the CHD5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CHD5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CHD5 locus and enabling the study of CHD5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CHD5 pathway restoration in tumor cells with silenced or reduced CHD5 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.